【目的】对1例中华蟾蜍多脏器寄生虫感染临床案例进行病理诊断并通过分子生物学鉴定其物种分类。【方法】对实验室饲养的1只消瘦、拒食后发生死亡的雌性中华蟾蜍进行解剖观察,病理组织学研究,采用蠕虫剖检法、结节压片法对分离获得的虫...【目的】对1例中华蟾蜍多脏器寄生虫感染临床案例进行病理诊断并通过分子生物学鉴定其物种分类。【方法】对实验室饲养的1只消瘦、拒食后发生死亡的雌性中华蟾蜍进行解剖观察,病理组织学研究,采用蠕虫剖检法、结节压片法对分离获得的虫体进行形态学观察研究,并进行分子生物学鉴定。【结果】剖检发现蟾蜍心脏、肝脏等脏器及多系膜寄生棘头虫,未在蟾蜍体内发现线虫、吸虫等其他蠕虫寄生。HE染色结果显示,棘头虫的感染寄生造成了蟾蜍肺脏、肝脏等多脏器的机械性损伤、炎性反应,其中肠壁黏膜发生严重损坏脱落。通过PCR扩增其细胞色素c氧化酶亚基Ⅰ(cytochrome c oxidase subunitⅠ,COⅠ)基因片段,获得5株不同部位的目的虫株COⅠ基因目的片段,经对比发现,鉴定的棘头虫均为同一种棘头虫,且其GC含量在40.14%~40.83%之间。选取JS-01株进行NCBI-BLAST比对,与似矛球吻棘头虫(Sphaerirostris lanceoide,登录号:MG931943.1)相似性最高,为99.5%。借助分子生物学分析并结合形态学观察结果确定其为似矛球吻棘头虫。【结论】似矛球吻棘头虫的大量感染寄生造成了蟾蜍的死亡,本试验结果可为药用蟾蜍棘头虫病的防控提供临床症状、病理、分子生物学等方面的参考,对两栖动物似矛球吻棘头虫的诊断与研究具有参考价值。展开更多
A total of 69 strains of thermophilic bacteria were isolated from water, soil and sediment samples from three Indonesia's hot spring areas (Pantai cermin, Kalianda and Banyu wedang) by using Minimal Synthetic Medi...A total of 69 strains of thermophilic bacteria were isolated from water, soil and sediment samples from three Indonesia's hot spring areas (Pantai cermin, Kalianda and Banyu wedang) by using Minimal Synthetic Medium (MSM). The extreme thermophile Brevibacillus sp. PLI-1 was found to produce extracellular thermophilic alkaline protease with optimal activity at 70℃ and pH 8.0-9.0. The molecular weight of the protease was estimated to be around 56 kD by SDS-PAGE. The maximum activity of the protease was 26.54 U mL-1. The protease activity did not decrease after 30 min and still retained more than 70% of relative activity after 60 min at 70℃ and pH 8.0. The ion Mg2+ was found to promote protease activity at both low and high concentrations, whereas Cu2+ and Zn2+ could almost completely inhibit the activity. Divalent cation chelator EDTA inhibited the enzyme activity by 55.06% ± 0.27%, while the inhibition caused by PMSF, Leupeptin, Pepstain A and Benzamidine were 66.78% ± 3.25%, 52.37% ± 0.25%, 62.47% ± 2.96% and 50.99% ± 0.24%, respectively. Based on these observations, the enzyme activity was conspicuously sensitive to the serine and cysteine protease inhibitors. All these results indicated that the protease isolated from the strain PLI-1 was a thermophilic protease and had a high-temperature stability and a pH stability.展开更多
CO2 fixation pathway of Caminibacter profundus, a chemolithoautotrophic e-Proteobacteria from deep-sea hydrothermal vent, was determined and characterized by genetic and enzymatic analyses. Gene expression of key enzy...CO2 fixation pathway of Caminibacter profundus, a chemolithoautotrophic e-Proteobacteria from deep-sea hydrothermal vent, was determined and characterized by genetic and enzymatic analyses. Gene expression of key enzymes for CO2 fixation in response to salinity, pH and O2 in Medium 829 were also investigated. The results demonstrate that C. profundus contained aclB, porA and oorA, the genes encoding key enzymes of reductive tricarboxylic acid （rTCA） cycle. However, genes fragments of cbbL and cbbMencoding key enzyme of Calvin cycle were not recovered. Key enzymatic activities of ATP citrate lyase （ACL）, pyruvate： ferredoxin oxidoreductase （POR） and 2-oxoglutarate： ferredoxin oxidoreductase （OOR） were also present in C. profun- dus. The combination of genetic and enzymatic analyses confirm that C. profundus adopted rTCA cycle for carbon assimilation. The results of aclB and oorA relative expressions of C. profundus demonstrate that the ranges of environmental factors for high genes expression were sea salt 3.0%-5.0% （optimum 3.0%）, pH 5.0-6.5（optimum pH 6.5）, anaerobic to microaerobic conditions （optimum 1.0% 02）. Gene expression pat- terns under different conditions show similar patterns with bacterial growth, revealing that key rTCA cycle genes provided molecular basis for bacterial growth and propagation. Our results suggest that C. profun- dus could regulate key genes of rTCA cycle for carbon assimilation and energy metabolism in response to environmental fluctuations in hydrothermal vent.展开更多
Due to morphological plasticity and paucity of diagnostic morphological characters, the taxonomy of Kap- paphycus gets more and more confused with the expanding of commercial cultivation. In this study, the phylogenet...Due to morphological plasticity and paucity of diagnostic morphological characters, the taxonomy of Kap- paphycus gets more and more confused with the expanding of commercial cultivation. In this study, the phylogenetic relationship of 13 strains of introduced Kappaphycus species in China was defined using DNA molecular markers, such as 18S rDNA, rbcL and cox2-cox3 spacer region. The resolutions obtained by three different molecular markers were compared： both cox2--cox3 spacer region and rbcL sequences are eligible in inter- species identification of Kappaphycus, whereas cox2-cox3 spacer region is more variable than rbcL sequence. There is several basepairs＇ discrepancy among 18S rDNA sequences, while it is 100% identical among both cox2-cox3 spacer region and rbcL sequences of the ten strains of K. alvarezii. We suppose that 18S rDNA sequence can provide more information in biogeography study of Kappaphycus than other two DNA sequences.展开更多
SNARE proteins are a group of membrane-associated proteins involved in exocytosis, secretion and membrane trafficking events in eukaryotic cells. Research on SNARE protein biology has become a more attractive field in...SNARE proteins are a group of membrane-associated proteins involved in exocytosis, secretion and membrane trafficking events in eukaryotic cells. Research on SNARE protein biology has become a more attractive field in recent years, which is applied to marine biology specifically to the fish Tilapia （Oreochromis niloticus）. Plasma membrane fractions of different tissues of Tilapia, including brain, liver-pancreas, intestine, skin and muscle, were extracted, and immuno-decorated with isoform-specific antibodies to the SNARE families and associated proteins. The presence of Syntaxins - 1 A, 2 and 3, SNAP - 23 and SNAP - 25, VAMP - 2, Munc - 18 - 1 and Munc - 13 in the brain was identified, which were differentially distributed in the other organ tissues of the fish Tilapia. The distinct distribution of SNARE and associated proteins will serve as the basis for further investigation into their special secretory function in these tissues of the fish.展开更多
In order to provide pertinent evidence of ion channel with immune response in the fish, whole cell patch-clamp technique was employed for potassium ion channel study in turbot （Scophthalmus maximus）. Lymphocytes wer...In order to provide pertinent evidence of ion channel with immune response in the fish, whole cell patch-clamp technique was employed for potassium ion channel study in turbot （Scophthalmus maximus）. Lymphocytes were isolated by Percoll density gradient centrifugation from peripheral blood samples, and electrophysiological characters of the channel were analyzed. In the recorded cells, activated voltage of the channels was -42.5±3.7 mV and the average peak current was 313.12±28.2 pA. The channel was identified as voltage dependent, the current was outward and it could be inhibited by 10 mmol/dma TEA or 5 mmol/dm^3 4-AP, a specific potassium channel inhibitor, identifying the existence of potassium channel in peripheral lymphocytes of the turbot.展开更多
Molecular genetic techniques have proven very useful for initial analysis of the extent of genetic variation and dispersal in several Antarctic moss species. In the present study, the small subunit ribosomal RNA （SSU...Molecular genetic techniques have proven very useful for initial analysis of the extent of genetic variation and dispersal in several Antarctic moss species. In the present study, the small subunit ribosomal RNA （SSU rDNA） and internal transcribed spacers of the nuclear ribosomal DNA （ITS rDNA） were sequenced in nine individuals of different mosses from the Fildes Peninsula of Antarctica. Sequence alignment showed that the extreme environment tended to increase the genetic diversity of Antarctic mosses. In addition, in our phylogenetic analysis, one previously unidentified Antarctic moss species was characterized by comparison with SSU and ITS rDNA sequences of known moss species. Moreover, the optimal culture medium and conditions for surface explant sterilization and protonemata induction in tissue culture of Pohlia nutans were investigated. The successful establishment of a tissue culture protocol together with the phylogenetic analysis of Antarctic mosses will provide technological support to establish an effective resource regeneration method for discovering new functional genes and gaining novel insights into the mechanisms of stress acclimation.展开更多
文摘【目的】对1例中华蟾蜍多脏器寄生虫感染临床案例进行病理诊断并通过分子生物学鉴定其物种分类。【方法】对实验室饲养的1只消瘦、拒食后发生死亡的雌性中华蟾蜍进行解剖观察,病理组织学研究,采用蠕虫剖检法、结节压片法对分离获得的虫体进行形态学观察研究,并进行分子生物学鉴定。【结果】剖检发现蟾蜍心脏、肝脏等脏器及多系膜寄生棘头虫,未在蟾蜍体内发现线虫、吸虫等其他蠕虫寄生。HE染色结果显示,棘头虫的感染寄生造成了蟾蜍肺脏、肝脏等多脏器的机械性损伤、炎性反应,其中肠壁黏膜发生严重损坏脱落。通过PCR扩增其细胞色素c氧化酶亚基Ⅰ(cytochrome c oxidase subunitⅠ,COⅠ)基因片段,获得5株不同部位的目的虫株COⅠ基因目的片段,经对比发现,鉴定的棘头虫均为同一种棘头虫,且其GC含量在40.14%~40.83%之间。选取JS-01株进行NCBI-BLAST比对,与似矛球吻棘头虫(Sphaerirostris lanceoide,登录号:MG931943.1)相似性最高,为99.5%。借助分子生物学分析并结合形态学观察结果确定其为似矛球吻棘头虫。【结论】似矛球吻棘头虫的大量感染寄生造成了蟾蜍的死亡,本试验结果可为药用蟾蜍棘头虫病的防控提供临床症状、病理、分子生物学等方面的参考,对两栖动物似矛球吻棘头虫的诊断与研究具有参考价值。
基金supported by the Basic Scientific Research Foundation of FIO, SOA (2010G13)the National Natural Science Foundation of China (41076108)+1 种基金the Science Foundation for Youth Scholars of State Oceanic Administration (2012131)the Public Welfare Project of Marine Science Research (200805039)
文摘A total of 69 strains of thermophilic bacteria were isolated from water, soil and sediment samples from three Indonesia's hot spring areas (Pantai cermin, Kalianda and Banyu wedang) by using Minimal Synthetic Medium (MSM). The extreme thermophile Brevibacillus sp. PLI-1 was found to produce extracellular thermophilic alkaline protease with optimal activity at 70℃ and pH 8.0-9.0. The molecular weight of the protease was estimated to be around 56 kD by SDS-PAGE. The maximum activity of the protease was 26.54 U mL-1. The protease activity did not decrease after 30 min and still retained more than 70% of relative activity after 60 min at 70℃ and pH 8.0. The ion Mg2+ was found to promote protease activity at both low and high concentrations, whereas Cu2+ and Zn2+ could almost completely inhibit the activity. Divalent cation chelator EDTA inhibited the enzyme activity by 55.06% ± 0.27%, while the inhibition caused by PMSF, Leupeptin, Pepstain A and Benzamidine were 66.78% ± 3.25%, 52.37% ± 0.25%, 62.47% ± 2.96% and 50.99% ± 0.24%, respectively. Based on these observations, the enzyme activity was conspicuously sensitive to the serine and cysteine protease inhibitors. All these results indicated that the protease isolated from the strain PLI-1 was a thermophilic protease and had a high-temperature stability and a pH stability.
基金China Ocean Mineral Resources R&D Association under contract No. DYXM-115-02-2-19Basic Scientific Fund for National Public Research Institutes of China under contract No. 2011T04
文摘CO2 fixation pathway of Caminibacter profundus, a chemolithoautotrophic e-Proteobacteria from deep-sea hydrothermal vent, was determined and characterized by genetic and enzymatic analyses. Gene expression of key enzymes for CO2 fixation in response to salinity, pH and O2 in Medium 829 were also investigated. The results demonstrate that C. profundus contained aclB, porA and oorA, the genes encoding key enzymes of reductive tricarboxylic acid （rTCA） cycle. However, genes fragments of cbbL and cbbMencoding key enzyme of Calvin cycle were not recovered. Key enzymatic activities of ATP citrate lyase （ACL）, pyruvate： ferredoxin oxidoreductase （POR） and 2-oxoglutarate： ferredoxin oxidoreductase （OOR） were also present in C. profun- dus. The combination of genetic and enzymatic analyses confirm that C. profundus adopted rTCA cycle for carbon assimilation. The results of aclB and oorA relative expressions of C. profundus demonstrate that the ranges of environmental factors for high genes expression were sea salt 3.0%-5.0% （optimum 3.0%）, pH 5.0-6.5（optimum pH 6.5）, anaerobic to microaerobic conditions （optimum 1.0% 02）. Gene expression pat- terns under different conditions show similar patterns with bacterial growth, revealing that key rTCA cycle genes provided molecular basis for bacterial growth and propagation. Our results suggest that C. profun- dus could regulate key genes of rTCA cycle for carbon assimilation and energy metabolism in response to environmental fluctuations in hydrothermal vent.
基金Basic Scientific Fund for National Public Research Institutes of China under contract No. 2008G19
文摘Due to morphological plasticity and paucity of diagnostic morphological characters, the taxonomy of Kap- paphycus gets more and more confused with the expanding of commercial cultivation. In this study, the phylogenetic relationship of 13 strains of introduced Kappaphycus species in China was defined using DNA molecular markers, such as 18S rDNA, rbcL and cox2-cox3 spacer region. The resolutions obtained by three different molecular markers were compared： both cox2--cox3 spacer region and rbcL sequences are eligible in inter- species identification of Kappaphycus, whereas cox2-cox3 spacer region is more variable than rbcL sequence. There is several basepairs＇ discrepancy among 18S rDNA sequences, while it is 100% identical among both cox2-cox3 spacer region and rbcL sequences of the ten strains of K. alvarezii. We suppose that 18S rDNA sequence can provide more information in biogeography study of Kappaphycus than other two DNA sequences.
基金This work was funded by the State"863"Tech Program of China under contract High No.2002AA629120the National Natural Science Foundation of China under contract No.40476060the Canadian Institute for Health Research under contract No.M0P-64465.
文摘SNARE proteins are a group of membrane-associated proteins involved in exocytosis, secretion and membrane trafficking events in eukaryotic cells. Research on SNARE protein biology has become a more attractive field in recent years, which is applied to marine biology specifically to the fish Tilapia （Oreochromis niloticus）. Plasma membrane fractions of different tissues of Tilapia, including brain, liver-pancreas, intestine, skin and muscle, were extracted, and immuno-decorated with isoform-specific antibodies to the SNARE families and associated proteins. The presence of Syntaxins - 1 A, 2 and 3, SNAP - 23 and SNAP - 25, VAMP - 2, Munc - 18 - 1 and Munc - 13 in the brain was identified, which were differentially distributed in the other organ tissues of the fish Tilapia. The distinct distribution of SNARE and associated proteins will serve as the basis for further investigation into their special secretory function in these tissues of the fish.
基金The National Natural Science Foundation of China under contract Nos 40476060 and 40876089the State Key Basic Research Program under contract No. 2006CB101805
文摘In order to provide pertinent evidence of ion channel with immune response in the fish, whole cell patch-clamp technique was employed for potassium ion channel study in turbot （Scophthalmus maximus）. Lymphocytes were isolated by Percoll density gradient centrifugation from peripheral blood samples, and electrophysiological characters of the channel were analyzed. In the recorded cells, activated voltage of the channels was -42.5±3.7 mV and the average peak current was 313.12±28.2 pA. The channel was identified as voltage dependent, the current was outward and it could be inhibited by 10 mmol/dma TEA or 5 mmol/dm^3 4-AP, a specific potassium channel inhibitor, identifying the existence of potassium channel in peripheral lymphocytes of the turbot.
基金financially supported by the National Natural Science Foundation of China(Grant nos.40906103 and 41206176)the Chinese Polar Environment Comprehensive Investigation & Assessment Programs(Grant nos.CHINARE2012-02-01 and CHINARE2012-01-06)the Ocean Public Welfare Scientific Research Project of China(Grant no.201005031)
文摘Molecular genetic techniques have proven very useful for initial analysis of the extent of genetic variation and dispersal in several Antarctic moss species. In the present study, the small subunit ribosomal RNA （SSU rDNA） and internal transcribed spacers of the nuclear ribosomal DNA （ITS rDNA） were sequenced in nine individuals of different mosses from the Fildes Peninsula of Antarctica. Sequence alignment showed that the extreme environment tended to increase the genetic diversity of Antarctic mosses. In addition, in our phylogenetic analysis, one previously unidentified Antarctic moss species was characterized by comparison with SSU and ITS rDNA sequences of known moss species. Moreover, the optimal culture medium and conditions for surface explant sterilization and protonemata induction in tissue culture of Pohlia nutans were investigated. The successful establishment of a tissue culture protocol together with the phylogenetic analysis of Antarctic mosses will provide technological support to establish an effective resource regeneration method for discovering new functional genes and gaining novel insights into the mechanisms of stress acclimation.
